Abstract
Objective
To explore the influence of ultra-low carbon dioxide partial pressure (pCO2) on the monoclonal antibody (mAb) N-glycosylation profile in Chinese hamster ovary (CHO) cell culture.
Results
In fed-batch bioreactor cultures, lowering the pCO2 in the medium (< 25 mmHg) via increasing headspace aeration decreased the cell viability and mAb production in CHO cells. Additionally, mAb galactosylation under low pCO2 was approximately 27.45 ± 2.13%, noticeably higher than that observed under normal pCO2 (21.36 ± 1.66%) at harvest. However, all of the relevant intracellular nucleotide sugar concentrations were dramatically decreased to approximately 50% of the levels found under normal pCO2 on day 7. Real-time PCR revealed that the upregulation of galactosylation-related glycosyltransferase genes and substrate transporter genes played a critical role in the improved galactosylation under the ultra-low pCO2 condition.
Conclusions
In the bioreactor culture processes, ultra-low pCO2 demonstrated a positive effect on mAb galactosylation.
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Acknowledgements
This work was supported by the National Natural Science Foundation of China (Nos. 21406066, 21106045), the National High Technology Research and Development Program of China (863 Program) (No. 2012AA02A303), the Fundamental Research Funds for the Central Universities (No. 22221818014), and the Development Funds of Shanghai Zhangjiang National Independent Innovation Demonstration Zone (No. ZJ2015-ZD-002).
Supporting information
Supplementary Table 1—Primers used in real-time PCR to quantify the expression levels of specific N-galactosylation-related genes.
Supplementary Fig. 1—The glycosylation profiles of 2-AB-labeled N-glycans separated using UPLC under normal pCO2 (0.05 vvm, red) and low pCO2 (0.4 vvm, black) on day 11. Symbols: square, N-acetylglucosamine; circle, mannose; rhombus, galactose; triangle, fucose; star, sialic acid.
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10529_2018_2586_MOESM1_ESM.tif
The glycosylation profiles of 2-AB-labeled N-glycans separated using UPLC under normal pCO2 (0.05 vvm, red) and low pCO2 (0.4 vvm, black) on day 11. Symbols: square, N-acetylglucosamine; circle, mannose; rhombus, galactose; triangle, fucose; star, sialic acid. Supplementary material 1 (TIFF 43375 kb)
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Wang, C., Wang, J., Chen, M. et al. Ultra-low carbon dioxide partial pressure improves the galactosylation of a monoclonal antibody produced in Chinese hamster ovary cells in a bioreactor. Biotechnol Lett 40, 1201–1208 (2018). https://doi.org/10.1007/s10529-018-2586-4
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DOI: https://doi.org/10.1007/s10529-018-2586-4